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First published online May 8, 2009; 10.1104/pp.109.137612

Plant Physiology 151:1087-1095 (2009)
© 2009 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES - FOCUS ISSUE

Site-Specific Integration of Transgenes in Soybean via Recombinase-Mediated DNA Cassette Exchange[OA]

Zhongsen Li*, Aiqiu Xing, Bryan P. Moon, Richard P. McCardell, Kelly Mills and S. Carl Falco

DuPont/Pioneer Crop Genetics, Experimental Station, Wilmington, Delaware 19880

A targeting method to insert genes at a previously characterized genetic locus to make plant transformation and transgene expression predictable is highly desirable for plant biotechnology. We report the successful targeting of transgenes to predefined soybean (Glycine max) genome sites using the yeast FLP-FRT recombination system. First, a target DNA containing a pair of incompatible FRT sites flanking a selection gene was introduced in soybean by standard biolistic transformation. Transgenic events containing a single copy of the target were retransformed with a donor DNA, which contained the same pair of FRT sites flanking a different selection gene, and a FLP expression DNA. Precise DNA cassette exchange was achieved between the target and donor DNA via recombinase-mediated cassette exchange, so that the donor DNA was introduced at the locus previously occupied by the target DNA. The introduced donor genes expressed normally and segregated according to Mendelian laws.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Zhongsen Li (zhongsen.li{at}cgr.dupont.com).

[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.109.137612

* Corresponding author; e-mail zhongsen.li{at}cgr.dupont.com.

Received February 24, 2009; accepted May 5, 2009; published May 8, 2009.

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